Objective To assess the application of ultrasonography in prenatal diagnosis of fetal choledochal cysts.Methods The clinical data of 25 fetuses,who were diagnosed as fetal choledocho cysts using ultrasonography and followed up,were retrospectively analyzed.Results Among 25 cases,24 cases were confirmed as congenital choledochal cysts after normal delivery or abortion,and 1 case of liver cyst was misdiagnosed.The 24 cases' prenatal ultrasound showed liver cystic space occupying wihich was connected to intrahepatic bile duct or gallbladder.The cysts wall was thin,and the colour Doppler flow imaging CDFI showed that blood vessels circled around the cyst.In 24 fetuses with prenatal choledochal cysts,21 were fullterm birth,3 were aborted including 1 case with other congenital abnormalities.The size of choledochal cysts was not changed in 5 cases,and was enlarged with the gestational ages in remaining 16 cases fetuses.The choledochal cysts did not affect fetal growth and development.The newborn infants with congenital choledochal cysts were treated surgically within 8 months after birth and all recovered well.Conclusion Fetal choledochal cysts have typical sonographic manifestations.Ultrasonography can be used for prenatal diagnosis of congenital choledochal cysts,which can be successfully treated by surgery early after birth.

Objective To investigate the effect of calcipotriol on melanin synthesis by human melanocytes and its possible action mechanism.Methods Primary melanocytes were cultured with various concentrations(10~(-5),10~(-6),10~(-7),10~(-8),10~(-9),10~(-10) mol/L)of calcipotriol for 24 or 48 hours.Subsequently,MTT assay,NaoH assay.Dopa-oxidase assay,Western blot and semiquantitative RT-PCR were used to measure the cell proliferation of,melanin synthesis by.tyrosinase activity,protein and mRNA expression levels in the melanocytes.respectively.Those untreated melanocytes served as the control.Results The calcipotriol between 10~(-9) and 10~(-5) mol/L had no significant effect on the proliferation of cultured melanocytes(P＞0.05).while that of 10~(-9) and 10~(-8) mol/L increased tyrosinase activity by 137%and 123%,and enhanced melanin synthesis by 40.63%and 18.75%,respectively,compamd with untreated melanocytes(both P＜0.05).Moreover,the tyrosinase protein level increased by 270.4%(P＜0.05)in melanocytes treated with calcipotriol at 10~(-9) mol/L for 24 hours.The strongest tyrosinase activity and melanin synthesis was observed in melanocytes treated with calcipotriol of 10~(-9) moI/L.Conclusions The proliferation of melanocytes is unaffected by calcipotriol at 10~(-9) to 10~(-5) mol/L,but it can elevate the expression of tyrosinase protein,enhance tyrosinase activity,and promote melanin synthesis in melanocytes.

Objective To study the effect of thyroid-stimulating hormone (TSH) on human vascular endothelial cells and smooth muscle cells and to explore the roles of TSH in the development of atherosclerosis.Methods Human vascular endothelial cells and smooth muscle cells were cultured in vitro.MTT method was used to assay the effect of TSH on cell viability.Real-time PCR was used to determine the levels of endothelial nitric oxide synthase (eNOS),prostacyclin(PGI2),endothelin-1 (ET-1),plasminagen activator inhibitor(PAI-1) (mRNA) in endothelial cells and the phenotype transition of smooth muscle cells.The effect of TSH to the cycle of smooth muscle cells was detected by using flow cytometry.Immunocytochemistry and Western blot were used to investigate the expression of the cell cyclin A,D1,and the expression of endothelial cell associated factors eNOS and ET-1.Results Compared with the control group,eNOS and PGI2 mRNA levels decreased while ET-1 and PAI-1 mRNA levels increased when different concentrations of TSH were applied to endothelial cells(P＜0.05).The level of eNOS protein was decreased gradually while the level of ET-1 protein was gradually increased(P＜0.05).Different concentrations of TSH applied to smooth muscle cells could promote the transition of cell cycle phase G2 to phase M and increase the expression of cell cyclin A and D1.Conclusion TSH may damage the function of vascular endothelial cells and promote the proliferation of smooth muscle cells.

This study was aimed to establish a method for the rapid content determination of Ixerin Z and 11,13α-dihydroixerin Z in Ku-Die-Zi (KDZ) injection by UPLC-ESI-MS/MS. The separation was performed on a Waters ACQUITY BEH C18 column (2.1 mm × 50 mm, 1.7μm) by using a gradient elution with the mobile phase of acetonitrile-water at the flow rate of 0.4 mL·min-1. The column temperature was set at 40℃. Multi-reaction moni-toring (MRM) scanning was employed for quantification in ESI negative mode. The results showed that two sesquit-erpene lactones in KDZ injection were totally separated within 2 min. The linear range of Ixerin Z was 5.70-182.50 ng·mL-1, and the linear range of 11,13α-dihydroixerin Z was 4.60-131.25 ng·mL-1. The correlation coefficient r was more than 0.999 0. The recovery rates (n = 6) were 98.18% and 97.52%, with RSDs < 1.5%. The established method was successfully applied for simultaneous content determination of Ixerin Z and 11,13α-dihydroixerin Z in 6 batches of KDZ injection from 2 factories, which had some variations on the content determination results. It was concluded that the method was rapid, accurate and sensitive, which can be used for the content determination of two sesquiterpene lactones in KDZ injection.

Objective To test the technical param eters of GlobalFiler?PC R A m plification K it for its ap-plication to forensic application value and to investigate the genetic polym orphism s. Methods The valida-tion w as conducted in sensitivity, m ixed sam ples, species specificity, adaptability, survivability, consistency, peak height balance and stability. The am plification and detection of the genom ic D N A from 373 unre-lated individuals from B eijing H an nationality w ere extracted by autom ation w orkstation. Results Global-Filer?PC R A m plification K it w as adaptive to som e m ixed, degraded and inhibited sam ples. The pow er of sensitivity and adaptability and peak height balance show ed w ell. The distributions of genotype fre-quencies for 21 STR loci in the population w ere all in accordance w ith H ardy-W einberg equilibrium (P>0.05). The PIC value of the 21 STR loci w as am ong 0.536 to 0.940; the H value w as am ong 0.558 to 0.933; the D P value w as am ong 0.783 to 0.992; the PE value w as am ong 0.243 to 0.874. Conclusion GlobalFiler?PC R A m plification K it is suitable for crim inal cases and D N A database in forensic practice. A nd 21 STR loci in B eijing H an nationality have high polym orphism , w hich have ap-plication value in forensic practice and population genetics.

OBJECTlVE To study the anti-HBV activity of prepared recombinant human serum aIbu-min-interferon α-2b fusion protein(HSA-IFNα-2b) in vitro. METHODS HepG2 ceIIs were infected with recombinant adenovirus with green fIuorescence protein and 1.6-foId HBV DNA(AdGFP-HBV). The ex-pression of HBV antigens,HBsAg and HBeAg in cuIture medium was detected by ELISA assay. The tox-icity of HSA-IFNα-2b on HepG2 ceIIs was evaIuated by mTT assay.The reIative expression of HBV RNA in ceIIs and the absoIute quantity of HBV DNA in cuIture supernatant were determined by quantitative PCR assay. The activity of HBV enhancer Ⅰ was detected by DuaI-Reporter gene assay. RESULTS HBV couId repIicate and express in HepG2 ceIIs after infection with AdGFP-HBV. The expression of HBsAg and HBeAg in cuIture serum of HepG2 ceIIs infected with AdGFP-HBV decreased by 51.32%(P﹤0.01)and 50.26%(P﹤0.01),respectiveIy,when HSA-IFNα-2b 500 kU·L-1 was added. The same concentration of HSA-IFNα-2b didn't inhibit the proIiferation of HepG2 ceIIs,but inhibited HBsAg in a concentration-dependent manner. The regression formuIa between HBsAg inhibitory rate(Y)and con-centration of HSA-IFNα-2b(X)was Y=21.11 IgX+11.91(r 2 = 0.954),IC50 = 63.76 kU·L-1 . HBV RNA in ceIIs and HBV DNA in the cuIture serum decreased by 52.83%(P﹤0.01)and 53.07%(P﹤0.01), respectiveIy,when HSA-IFNα-2b 500 kU·L-1 was added. The activity of enhancer Ⅰ decreased by 40.04%(P﹤0.01)when HSA-IFNα-2b 500 kU·L-1 was added. CONCLUSlON The ceII modeI of HBV repIication for evaIuating anti-HBV agents is successfuIIy estabIished. HSA-IFNα-2b exhibits noticeabIe anti-HBV effect invitro.

Objective To study the plasma D-dimer (DD) level changes in the patients with rheumatoid arthritis (RA), and to analyze its correlation with various clinical indicators. Methods Selected a total of 138 cases of RA patients and 50 cases of normal person as matched group the plasma D-dimer level and the correlation analysis with age, rheuma-toid factor (RF), erythrocyte sedimentation rate (ESR), visual analogue scale (VAS score)and platelet were analyzed. Results The level of D-dimer level in RA patients was significantly higher than that of healthy controls, and the dif-ference was statistically significant (P<0.01);The D-dimer level of high disease activity group and low disease activity group with RA patients were significantly higher than the remission groups of RA patients (P all <0.01); The D-dimer level in high disease activity group of RA patients was significantly higher than the low disease activity group (P<0.01); There were positive correlations between the D-dimer level and the age, ESR, VAS score, RF, platelet in all RA patients (P all <0.01). Conclusion There is high coagulation state in RA patients, so the D-dimer can be used as a clinical nonspecific inflammatory reaction index in patients with RA, and can direct the clinical treatment.

OBJECTIVETo construct a dual-reporter gene system that will be applicable for use as a tool to screen and evaluate therapeutic drug compounds that inhibit transcription of the gene encoding collagen I, chain at1 (COL1A1).

METHODSThe full-length eDNA of transforming growth factor beta1 (TGFbeta1) was cloned by RT-PCR and inserted into two vectors, pcDNA3.1 and pJW4303, for construction of two eukaryotic expression vectors, pcDNA3.1-TGFbeta1 and pJW4303-TGFbeta1.Next, the promoter region of COL1A1, cloned by PCR using human genome DNA as template, was inserted into the vector pGL4.29 to construct the reporter gene vector, pGL4.29-COL1A1 promoter.All three recombinant vectors were verified by restriction enzyme digestion and DNA sequencing.Either the pcDNA3.1-TGFbeta1 or pJW4303-TGFbeta1 vector along with the pGL4.29-COL1A1 promoter vector or a pRL-null, control reporter, vector were co-transfected into the LX-2 human hepatic stellate cells to establish the transcription-activated dualreporter gene system.This system was used as a cell model for screening anti-liver fibrosis compounds that inhibit the transcription of COL1A1.Dexamethasone, a model drug that is known to inhibit the expression of COL1A1, was used as a control to validate the dual-reporter gene system.

RESULTSThe two TGFbeta1-expressing vectors and the reporter gene vector containing the promoter region of COL1A1 were successfully constructed.The results of a dual-reporter gene assay showed that TGFbeta1 co-expression increased the activity of the COL1A1 promoter by above 200-fold (t =21.78, P =0.0001), whereas in the absence of TGF31 co-expression the activity was below 2-fold (t =3.396, P =0.0274).The transcriptionactivated dual-reporter gene system was successfully established.The model drug, dexamethasone, effectively inhibited the activity of the COL 1A1 promoter in dose-dependent manner; the activity decreased 29.6% with 10 mumol/L dexamethasone (t =4.140, P =0.0144) and 53.9% with 100 mumol/L (t =6.193, P =0.0035).

CONCLUSIONThe dual-luciferase reporter system of TGFbeta1 and COL1A1 co-expression developed here can be used as a cell model to screen and evaluate anti-liver fibrosis compounds that inhibit activity of the COL1A1.

Base Sequence ; Collagen Type I ; genetics ; Drug Evaluation, Preclinical ; Genes, Reporter ; Genetic Vectors ; Humans ; Liver Cirrhosis ; drug therapy ; Luciferases ; Promoter Regions, Genetic ; Transcriptional Activation ; drug effects ; Transfection ; Transforming Growth Factor beta1

Objectives To conduct a research on the possibility and effect factors of latent fingerprints development in clothing objects after vacuum coating, and extracting fingerprints DNA and to probe in the relation among DNA template quantity and genetic loci numbers tested, and the rfu value after coating. Methods To select two groups that are free sweat hands and sweat hands and have them press their fingerprints on the cloth, after coating, and to analyze the effect of time, to quantify and test the targeted fingerprints DNA, to compare the locus numbers tested between white and black cloth. Results As the time is prolonged, the locus numbers tested decrease. The locus numbers tested on the group of sweat hands using the same method after the same placed time are lager than the free sweat hands. When the value of rfu is 600 above, the ratio of the locus numbers tested is more than 90% and the threshold of templates is 0.013ng. The locus numbers tested of white cloth is larger, comparing with black cloth when using the same method. What is more, there exists an prohibitive influence of pigments of the dyed cloth over the PCR amplification, to put it further, the loci numbers tested will be trimmed. Conclusion The technology of vacuum coating can be well used in the area of detecting fingerprint DNA.

Objective To explore the spatial and temporal characteristics of human brucellosis in Shandong province and to provide scientific basis for the development of related regional public health strategies.Methods 1 802 diagnosed cases of human brucellosis patients were selected based on the data that was collected by Diseases Reporting Information System between year 2004 and 2012 in Shandong province.Methods on spatial thematic mapping,spatial autocorrelation analysis,spatial clustering analysis,and temporal clustering analysis were applied to describe the temporal and spatial distribution on human brucellosis cases.Results The incidence rate of human brucellosis increased from 0.038 2/100 000 (35 cases) to 0.620 5/100 000 (598 cases),with annual average incidence rate as 0.211 1/100 000 and the incidence was evidently increased.The value of M (0.375 3) showed that this disease was seasonal,with the epidemic months between March and June,accounting for 56.27％ (1 014/1 802).The Global Moran' s I index was 0.198 901 (P=0.000 120),showing that there was a positive correlation between space and the incidence of brucellosis.The incidence rates in 2006,2007,2009 and 2012 and the space distribution appeared a positive correlation (P＜0.05) in Shandong province.The local Moran's I index showed that there were 8 “High-High” (HH) clustering areas,which were proved to have statistical significance (P＜0.05).Local indicators of spatial association (LISA) revealed that southwest and north districts of Shandong were highly clustered districts of brucellosis and the areas paralleled to the areas that having higher incidence rates.There were two spatial clustering areas in this study,one as the center of Juanchen with radiation radius at 33.83 km whose RR was 9.78 (P＜0.05) and the other was the center of Binchen with radiation radius at 62.78 km with RR as 4.99 (P＜0.05).All the 8 HH counties (districts) were included in the two cluster regions.Conclusion Incidence of human brucellosis showed an obvious increase in Shandong during year 2004-2012.Months with epidemics were between March and June.The incidence of brucellosis in counties (districts) was non-randomly distributed.A positive spatial correlation and the feature of clusters was noticed.